Vikas K Singh

5 Questions 9 Answers 0 Followers

Questions related from Vikas K Singh

Is it possible do ammonium sulphate fractionation/precipitation of a protein inclusion bodies (IB's) dissolved in 8M Urea?

My protein (64kDa) expressed in insoluble inclusion bodies. I tried to purify it by ion-exchange and affinity but unable to purify protein. So, want to try ammonium sulfate precipitation of...

07 March 2018 670 4 View

What could be the reason for E.coli BL21 (DE3) cells turning into pink and green color?

Upon induction with IPTG in TB media, cells turned into pinkish color and yielded brownish inclusion body; while same BL21 culture inoculated in LB media resulting in green color. I have been...

07 June 2017 2,048 5 View

How to make competent BL21 (DE3) cell ? Is there any specific precaution needs to be taken care of?

I am unable to make BL21 DE3 competent cells. I have tried many protocols including CaCl2, MgCl2, and TSS method but every time I failed. On the other hand, if I prepare competent cells for DH5a...

30 April 2017 4,097 2 View

What cause up shift of protein band in SDS-PAGE?

I am working on recombinant protein (Ig's) expressed in form of inclusion bodies. Upon giving certain treatments there is a up shift in protein band. If my protein is degrading during the process;...

20 November 2016 1,869 3 View

What is the concentration of dNTP's inside the cell during replication?

As we know final concentration of dNTP's in PCR reaction is 0.2mM and during replication intracellular concentration of rNTPs is 10–100-fold or more higher than that of dNTPs. But what is the...

11 March 2016 1,458 1 View